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Immune signatures of protective spleen memory CD8 T cells

View Article: PubMed Central - PubMed

ABSTRACT

Memory CD8 T lymphocyte populations are remarkably heterogeneous and differ in their ability to protect the host. In order to identify the whole range of qualities uniquely associated with protective memory cells we compared the gene expression signatures of two qualities of memory CD8 T cells sharing the same antigenic-specificity: protective (Influenza-induced, Flu-TM) and non-protective (peptide-induced, TIM) spleen memory CD8 T cells. Although Flu-TM and TIM express classical phenotypic memory markers and are polyfunctional, only Flu-TM protects against a lethal viral challenge. Protective memory CD8 T cells express a unique set of genes involved in migration and survival that correlate with their unique capacity to rapidly migrate within the infected lung parenchyma in response to influenza infection. We also enlighten a new set of poised genes expressed by protective cells that is strongly enriched in cytokines and chemokines such as Ccl1, Ccl9 and Gm-csf. CCL1 and GM-CSF genes are also poised in human memory CD8 T cells. These immune signatures are also induced by two other pathogens (vaccinia virus and Listeria monocytogenes). The immune signatures associated with immune protection were identified on circulating cells, i.e. those that are easily accessible for immuno-monitoring and could help predict vaccines efficacy.

No MeSH data available.


Poised cytokines produced by Flu-TM and human memory CD8 T cells.(A) Cytokine profile of stimulated naive, TIM or Flu-TM CD8 T cells. Supernatants were collected after 48 hours of NP68 peptide stimulation for indicated cell subsets. Data are mean ± SD of 3 independent experiments with a pool of four or five mice per group. (B) Cytokine production by sorted human naive or memory CD8 T cells. Supernatants were collected after 48 hours of anti-CD3/CD28 stimulation or anti-CD3/CD28 stimulation plus IL-12/IL-18 stimulation. Data are mean ± SD of 5 healthy donors. *p < 0.05 (two tailed unpaired t-test).
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f5: Poised cytokines produced by Flu-TM and human memory CD8 T cells.(A) Cytokine profile of stimulated naive, TIM or Flu-TM CD8 T cells. Supernatants were collected after 48 hours of NP68 peptide stimulation for indicated cell subsets. Data are mean ± SD of 3 independent experiments with a pool of four or five mice per group. (B) Cytokine production by sorted human naive or memory CD8 T cells. Supernatants were collected after 48 hours of anti-CD3/CD28 stimulation or anti-CD3/CD28 stimulation plus IL-12/IL-18 stimulation. Data are mean ± SD of 5 healthy donors. *p < 0.05 (two tailed unpaired t-test).

Mentions: The poised genes signature HP0RP1 associated with protective Flu-TM is strongly enriched in cytokines (Csf2 also known as Gm-csf, Il21 and Il10) and CC-chemokines (Ccl1 and Ccl9). Expression of these cytokines by memory CD8 T cells was validated at the mRNA level by PCR (Supplementary Fig. S4). The production of the poised RP1 cytokine GM-CSF and chemokines CCL1 and CCL9 was measured at the protein level following peptide stimulation. Results in Fig. 5A indicate that all three proteins are secreted in significantly larger quantities by Flu-TM compared to TIM or naive F5 cells. Another characteristic of memory CD8 T cells is their ability to increase their functional potential and secrete certain cytokines such as IFN-γ in response to two cytokines, IL-12 and IL-18, associated with inflammatory milieu36. Similarly, the production of GM-CSF by Flu-TM, but not CCL1 or CCL9, was strongly induced by IL-12 and IL-18 (Supplementary Fig. S5), indicating that its production by memory CD8 T cells follows the same pattern as IFN-γ. We next looked if the genes coding for these proteins were also behaving as poised genes in human memory CD8 cells. To that aim, we purified human blood memory CD8 T cells and measured their capacity to produce those cytokines/chemokines following stimulation with anti-CD3/CD28, in the presence or absence of IL-12/IL-18. Of note, CCL9 is not conserved in humans, so we focused our analysis on CCL1 and GM-CSF or IFN-γ as a control (Fig. 5B). As described before, GM-CSF or IFN-γ could readily be detected following stimulation8. We next could show that memory phenotype but not naive CD8 T cells also produced CCL1 chemokine, indicating that the CCL1 gene is also poised in human memory CD8 T cells.


Immune signatures of protective spleen memory CD8 T cells
Poised cytokines produced by Flu-TM and human memory CD8 T cells.(A) Cytokine profile of stimulated naive, TIM or Flu-TM CD8 T cells. Supernatants were collected after 48 hours of NP68 peptide stimulation for indicated cell subsets. Data are mean ± SD of 3 independent experiments with a pool of four or five mice per group. (B) Cytokine production by sorted human naive or memory CD8 T cells. Supernatants were collected after 48 hours of anti-CD3/CD28 stimulation or anti-CD3/CD28 stimulation plus IL-12/IL-18 stimulation. Data are mean ± SD of 5 healthy donors. *p < 0.05 (two tailed unpaired t-test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5121635&req=5

f5: Poised cytokines produced by Flu-TM and human memory CD8 T cells.(A) Cytokine profile of stimulated naive, TIM or Flu-TM CD8 T cells. Supernatants were collected after 48 hours of NP68 peptide stimulation for indicated cell subsets. Data are mean ± SD of 3 independent experiments with a pool of four or five mice per group. (B) Cytokine production by sorted human naive or memory CD8 T cells. Supernatants were collected after 48 hours of anti-CD3/CD28 stimulation or anti-CD3/CD28 stimulation plus IL-12/IL-18 stimulation. Data are mean ± SD of 5 healthy donors. *p < 0.05 (two tailed unpaired t-test).
Mentions: The poised genes signature HP0RP1 associated with protective Flu-TM is strongly enriched in cytokines (Csf2 also known as Gm-csf, Il21 and Il10) and CC-chemokines (Ccl1 and Ccl9). Expression of these cytokines by memory CD8 T cells was validated at the mRNA level by PCR (Supplementary Fig. S4). The production of the poised RP1 cytokine GM-CSF and chemokines CCL1 and CCL9 was measured at the protein level following peptide stimulation. Results in Fig. 5A indicate that all three proteins are secreted in significantly larger quantities by Flu-TM compared to TIM or naive F5 cells. Another characteristic of memory CD8 T cells is their ability to increase their functional potential and secrete certain cytokines such as IFN-γ in response to two cytokines, IL-12 and IL-18, associated with inflammatory milieu36. Similarly, the production of GM-CSF by Flu-TM, but not CCL1 or CCL9, was strongly induced by IL-12 and IL-18 (Supplementary Fig. S5), indicating that its production by memory CD8 T cells follows the same pattern as IFN-γ. We next looked if the genes coding for these proteins were also behaving as poised genes in human memory CD8 cells. To that aim, we purified human blood memory CD8 T cells and measured their capacity to produce those cytokines/chemokines following stimulation with anti-CD3/CD28, in the presence or absence of IL-12/IL-18. Of note, CCL9 is not conserved in humans, so we focused our analysis on CCL1 and GM-CSF or IFN-γ as a control (Fig. 5B). As described before, GM-CSF or IFN-γ could readily be detected following stimulation8. We next could show that memory phenotype but not naive CD8 T cells also produced CCL1 chemokine, indicating that the CCL1 gene is also poised in human memory CD8 T cells.

View Article: PubMed Central - PubMed

ABSTRACT

Memory CD8 T lymphocyte populations are remarkably heterogeneous and differ in their ability to protect the host. In order to identify the whole range of qualities uniquely associated with protective memory cells we compared the gene expression signatures of two qualities of memory CD8 T cells sharing the same antigenic-specificity: protective (Influenza-induced, Flu-TM) and non-protective (peptide-induced, TIM) spleen memory CD8 T cells. Although Flu-TM and TIM express classical phenotypic memory markers and are polyfunctional, only Flu-TM protects against a lethal viral challenge. Protective memory CD8 T cells express a unique set of genes involved in migration and survival that correlate with their unique capacity to rapidly migrate within the infected lung parenchyma in response to influenza infection. We also enlighten a new set of poised genes expressed by protective cells that is strongly enriched in cytokines and chemokines such as Ccl1, Ccl9 and Gm-csf. CCL1 and GM-CSF genes are also poised in human memory CD8 T cells. These immune signatures are also induced by two other pathogens (vaccinia virus and Listeria monocytogenes). The immune signatures associated with immune protection were identified on circulating cells, i.e. those that are easily accessible for immuno-monitoring and could help predict vaccines efficacy.

No MeSH data available.